• 한국가금학회:학술대회논문집
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• 한국가금학회 2002년도 가을 학술발표논문집
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• pp.116-117
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• 2002

보존온도별(5, 25, 35$^{\circ}C$)로 정액과 희석액의 비율(1：1, 1：2, 1：3)을 달리하여 보존시간(1, 3, 6시간)에 따른 정자운동성을 조사한바, 5$^{\circ}C$ 냉장 온도에서는 희석배율에 따른 보존시간별 정자운동성은 차이가 없었으나, $25^{\circ}C$ 상온에서 6시간, 35$^{\circ}C$ 고온에서 3시간 이상 보존할 때 정액과 희석액의 비율이 1：1보다는 1：2 비율에서 정자운동성이 현저히 높게 유지되었다(P<0.05). 온도가 높을수록 1：1 배율보다는 1：2 이상 희석배율을 높여 주었을 때 정자의 운동성을 높게 유지시킬 수 있었다. Skim milk glucose액을 사용하여 닭 정액을 희석한 후 인공수정 하였을 때, 1회 주입정자수를 0.2, 0.4, 1 및 2억으로 하였을 때 각각 90.67, 94.00, 96.00 및 98.67%의 수정율을 나타냈다. 90%이상의 수정율을 얻기 위해서는 0.2억 이상의 정자 주입이 필요하며, 94%이상 안정적인 수정율을 얻기 위해서는 0.4억의 정자가 필요하다고 판단된다.

• Journal of Animal Science and Technology
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• 제48권6호
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• pp.777-784
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• 2006

본 연구는 돼지에서 정자 SCSA와 정자 운동성 및 인공수정을 이용한 분만율과의 상관관계를 조사하기 위해 인공수정센터에서 정액 생산용으로 이용되고 있는 종모돈 중 개체별 정액을 이용한 인공수정 번식성적을 가지고 있는 종모돈 26두의 정액 성상 기록과 번식성적을 분석 하였고, 액상정액을 공시하여 정자운동성 분석과 SCSA를 실시하였다. 종모돈의 정액 채취 횟수에 따른 SCSA 분석은 종모돈 26두를 공시하여 정액을 8주간 6회 채취하여 공시하였다. 종모돈은 인공수정 분만율(80% 이상 ; 종모돈 7두, AI 256복, 8070% ; 종모돈 9두, AI 745복 및 70% 미만 ; 종모돈 10두, AI 293 복)에 따라 3개 군으로 구분 하였다. 액상 정액의 CASA 분석 결과는 분만율에 따른 종모돈 군 간에 유의적인 차이는 인정되지 않았다. 종모돈의 SCSA 결과는 분만율이 높을수록 COMP αt, %Red, %Peak R 및 %Mean R 값은 낮게 나타났고, 분만율이 가장 낮은 종모돈 군과 다른 두 군간에 유의적인 차이를 보였다(P<0.05). 종모돈 정액을 채취 차수에 따른 정액의 성상과 SCSA를 조사한 결과에서 채취 차수에 따른 차이는 나타나지 않았다. 종모돈의 인공수정 분만율과 SCSA 결과와의 상관관계는 COMP αt, SD αt, %Red, %Peak R, Mean R는 모두 부의 상관관계를 나타냈다. 이상의 결과에서 SCSA는 종모돈 정액의 수정 능력을 평가하고 예측하는데 유용한 방법으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제22권2호
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• pp.131-141
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• 1995

This study was designed to determine the relationship between sperm acrosome reaction following ionophore challenge(ARIC) and hamster ovum sperm penetration assay(SPA) as assessment of fertilizing capacity of male. ARIC test and SPA were performed in 23 fertile and 19 subfertile men. The results were as follows; Sperm concentration was significantly higher in fertile group compared with subfertile group: $114.6{\pm}64.40$ vs $61.3{\pm}46.50{\times}10^6/ml$. However, there were no significantly differences in seminal volume, motility and motility index, respectively. There was a significantly correlation between spontaneous and induced AR in fertile and subfertile group, respectively. ARIC value was significantly higher in fertile group, compared with subfertile group: $12.0{\pm}5.57%$ vs $2.6{\pm}4.96%$. Both Penetration rate(PR) and Penetration index(PI) were significantly higher in fertile group, compared with subfertile group: $97.4{\pm}7.40%$ vs $64.9{\pm}36$. 20% and $5.4{\pm}2.88$ vs $1.5{\pm}1.47$, respectively. The Positive predictive value(PPV), Negative predictive value(NPV), sensitivity and specificity of ARIC test (cut-off: 8.5) and SPA(PI cut-off : 3.0) in predicting fertility were 95.0%, 81.8%, 82.6%, 94.7% and 95.2%, 85.7%, 87.0% and 94.7%, respectively. There was no significantly difference in predicting fertility between ARIC test and SPA. In conclusion, ARIC test was shown to have a predictive value for fertilizing capacity comparable to that of the hamster ovum sperm penetration assay. Therefore, ARIC test may be a simple and cost-effective addition to existing semenology instead of SPA.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.26-26
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• 2003

Hyaluronic acid (HA)-binding sites have been shown the diagnostic potential fur assessment of sperm maturity, which is related to male fertility. This study was designed to evaluate chromosomal patterns in porcine embryos produced by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with non- or HA-binding sperm (HABS). For binding of sperm with HA, sperm incubated in 10 ${mu}ell$ drop containing HA (0.8 mg/ml)-agarose (0.8%) mixture for 15 min. IVF and ICSI with non- or HA-bound sperm examined with matured oocytes at 44 hr after in vitro maturation. Embryos were cultured in 50 ${mu}ell$ of NCSU 23 containing 0.5% BSA for 5 days and then in 50 ${mu}ell$ of NCSU 23 containing 10% FBS for 2 days. For the evaluation of chromosomal aneuploidies, chromosome 1 sub-metacentric specific probe was used in sperm and embryos by fluorescence in situ hybridization (FISH). The frequency of aneuploidy sperm for chromosome 1 was 6.25%. The significant differences following IVF and ICSI with non- or HA-bound sperm were not observed in blastocyst formation rates (18.6, 23.5, and 23.8%) and cell number (61.8 $\pm$ 12.5, 55.5 $\pm$ 7.3, and 59.3 $\pm$ 9.6). Moreover, the percentage of diploidy in 4-cell stage embryos was 57.1% (IVF), 68.8% (ICSI), and 76.3% (ICSI-HABS). These results suggest that HA-binding sites may be a material for selection of normal sperm for ICSI. Therefore HA selection of normal sperm may be reduce the loss to embryonic mortality prior to embryo transfer in pig.

• 한국양식학회지
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• 제12권1호
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• pp.57-62
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• 1999

숭어 정액의 냉장 및 냉동보존시 적합한 희석액을 선정하고, 알에 대한 수정률을 평가하였다. 비보존 숭어 정자의 머리는 공모양으로 직경 $1.26{\pm}0.08 \{mu}textrm{m}$, 길이 $1.06{\pm}0.07 \{mu}textrm{m}$ 였으며, 과립상의 염색질을 가지고 있었다. 숭어 정자의 냉장보존시($0^{\circ}C$, 10일간) 희석액으로는 동종의 혈정이 가장 높은 정자활성을 나타냈으며, egg-tris, 0.1M, 0.3M 및 0.5M glucos에서는 활성이 서로 비슷하였다. 또한 동해방지제로 10% DMSO, 희석액으로 MFRS를 사용하여 냉동보존한 후 해동시켰을 때 대조구와 유사한 수정률을 보였다. 냉동보존 후 해동시킨 일부 정자 중에서는 세포막이 이탈되거나 소실되는 구조적 변화를 나타냈다.

• 한국수정란이식학회지
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• 제32권1호
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• pp.9-15
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• 2017

The present study was aimed to determine the effect of green tea extract (GTE) and beta-mercaptoethanol (${\beta}$-ME) supplementation in boar sperm freezing extender on sperm motility, viability and reactive oxygen species (ROS) level. Experimental groups were allocated into Lactose-egg yolk (LEY) without antioxidant (control), GTE (1,000 mg/L GTE in LEY) and ${\beta}$-ME ($50{\mu}M$ ${\beta}$-ME in LEY). Spermatozoa extended with LEY were cooled to $5^{\circ}C$ for 3 h and then kept at $5^{\circ}C$ for 30 min following dilution with LEY containing 9% glycerol and 1.5% Equex STM (final sperm concentration: $1{\times}10^8/mL$). Spermatozoa were loaded into straws and frozen in nitrogen vapor for 20 min. Following thawing at $37^{\circ}C$ for 25 sec, sperm viability and ROS level were measured using fluorescent double stain Fertility(R) and cytometry, respectively. Motility and viability of GTE supplemented-group were higher than those of control and ${\beta}$-ME without significance. ROS level in GTE group showed significantly lower than control (P < 0.05). In conclusion, GTE supplementation in boar sperm freezing extender can reduce ROS generation during freezing.

• Toxicological Research
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• 제18권2호
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• pp.167-174
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• 2002

Historical control data have been shown to be valuable in the proper interpretation and validation of reproductive toxicology studies. The present data were compiled from rat fertility and early embryonic development studies conducted at Korea Institute of Toxicology during the 1994∼2001 period. These data were assembled in order to provide background information for the general and reproductive data collected in 11 fertility and early embryonic development studies using Sprague-Dawley rats obtain-ing from the Breeding Facility, Korea Institute of Toxicology, Korea. A total of 274 males and 274 females were used in these studies during the eight-year period. Parameters of fertility and early embryonic development included clinical sign, body weights, food consumption, organ weights, estrus cycle, copulation index, precoital time, fertility index, pregnancy index, sperm parameters, and early embryonic development parameters. Most of the values were comparable to the previous historical control data reported by other investigators. These data can be wed not only as a historical data base for the meaningful interpretation of data from reproductive and developmental toxicity studies, but also as a contribution to biological characterization of Sprague-Dawley rats.

• Asian-Australasian Journal of Animal Sciences
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• 제33권9호
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• pp.1411-1420
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• 2020

Objective: The aim of study was to investigate the effects of season and single layer centrifugation (SLC) before cryopreservation on post-thaw bull sperm quality in Thailand. Methods: Semen was collected from 6 bulls (Bos indicus) in summer, rainy season and winter 2014 through 2016. Semen characteristics, sperm morphology, sperm kinematics, viability, chromatin structure and mitochondrial membrane were evaluated. Meteorological data were available from the local meteorological station; Results: Season had an effect on semen characteristics in the raw ejaculate, with higher proportions of normal spermatozoa and lower abnormalities in winter than in the other two seasons. Sperm kinematics, viability, DNA fragmentation index, and mitochondrial membrane potential were not different between seasons. Sperm samples selected by SLC had greater normal morphology and a lower proportion with bent tails than controls and higher values of progressive motility (PRO), beat cross frequency, linearity, straightness, wobble (WOB), and lower values of slow motility, velocity average path (VAP), velocity curved line, and amplitude of lateral head displacement than controls. In addition, SLC-selection had a favorable effect on PRO, VAP, and WOB that differed among seasons. Conclusion: Our results suggested that these bulls were well adapted to their location, with season having an effect on sperm morphology. Moreover, SLC could be used prior to cryopreservation, regardless of season, to enhance normal morphology and kinematics of bull sperm samples without adversely affecting other parameters of sperm quality. However, there was considerable variation among bulls in DNA fragmentation index, mitochondrial membrane potential and sperm viability. In addition, SLC had a positive effect on sperm morphology and sperm kinematics, which could be expected to influence fertility.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2000년도 제39차 춘계 학술대회
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• pp.13-28
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• 2000

Human spermatozoa exhibit a capacity to generate ROS and initiate peroxidation of the unsaturated fatty acids in the sperm plasma membrane, which plays a key role in the etiology of male infertility. The short half-life and limited diffusion of these molecules is consistent with their physiologic role in key biological events such as acrosome reaction and hyperactivation. The intrinsic reactivity of these metabolites in peroxidative damage induced by ROS, particularly $H_2O_2$ and the superoxide anion, has been proposed as a major cause of defective sperm function in cases of male infertility. The number of antioxidants known to attack different stages of peroxidative damage is growing, and it will be of interest to compare alpha-tocopherol and ascorbic acid with these for their therapeutic potential in vitro and in vivo. Both spermatozoa and leukocytes generate ROS, although leukocytes produce much higher levels. The clinical significance of leukocyte presence in semen is controversial. Seminal plasma confers some protection against ROS damage because it contains enzymes that scavenge ROS, such as catalase and superoxide dismutase. A variety of defense mechanisms comprising a number of antioxidants can be employed to reduce or overcome oxidative stress caused by excessive ROS. Determination of male infertility etiology is important, as it will help us develop effective therapies to overcome excessive ROS generation. ROS can have both beneficial and detrimental effects on the spermatozoa and the balancing between the amounts of ROS produced and the amounts scavenged at any moment will determine whether a given sperm function will be promoted or jeopardized. Accurate assessment of ROS levels and, subsequently, OS is Vital, as this will help clinicians both elucidate the fertility status and identify the subgroups of patients that respond or do not respond to these therapeutic strategies. The overt commercial claims of antioxidant benefits and supplements for fertility purposes must be cautiously looked into, until proper multicentered clinical trials are studied. From the current data it appears that no Single adjuvant will be able to enhance the fertilizing capacity of sperm in infertile men, and a combination of the possible strategies that are not toxic at the dosage used would be a feasible approach.

• The Korean Journal of Physiology and Pharmacology
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• 제23권5호
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• pp.381-392
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• 2019

Sperm function and male fertility are closely related to pH dependent $K^+$ current (KSper) in human sperm, which is most likely composed of Slo3 and its auxiliary subunit leucine-rich repeat-containing protein 52 (LRRC52). Onion peel extract (OPE) and its major active ingredient quercetin are widely used as fertility enhancers; however, the effect of OPE and quercetin on Slo3 has not been elucidated. The purpose of this study is to investigate the effect of quercetin on human Slo3 channels. Human Slo3 and LRRC52 were co-transfected into HEK293 cells and pharmacological properties were studied with the whole cell patch clamp technique. We successfully expressed and measured pH sensitive and calcium insensitive Slo3 currents in HEK293 cells. We found that OPE and its key ingredient quercetin inhibit Slo3 currents. Inhibition by quercetin is dose dependent and this degree of inhibition decreases with elevating internal alkalization and internal free calcium concentrations. Functional moieties in the quercetin polyphenolic ring govern the degree of inhibition of Slo3 by quercetin, and the composition of such functional moieties are sensitive to the pH of the medium. These results suggest that quercetin inhibits Slo3 in a pH and calcium dependent manner. Therefore, we surmise that quercetin induced depolarization in spermatozoa may enhance the voltage gated proton channel (Hv1), and activate non-selective cation channels of sperm (CatSper) dependent calcium influx to trigger sperm capacitation and acrosome reaction.